coomassie brilliant blue r  (Millipore)


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    Structured Review

    Millipore coomassie brilliant blue r
    Coomassie Brilliant Blue R, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/coomassie brilliant blue r/product/Millipore
    Average 86 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    coomassie brilliant blue r - by Bioz Stars, 2020-04
    86/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi
    Article Snippet: Total B. burgdorferi proteins were prepared by growing cells to stationary phase, harvesting cells by centrifugation, and washing cells twice in phosphate-buffered saline. .. Proteins were stained with Coomassie blue R-250 (Sigma).

    Whole Genome Amplification:

    Article Title: Enzymatic and Pro-Inflammatory Activities of Bothrops lanceolatus Venom: Relevance for Envenomation
    Article Snippet: .. Chemicals and Reagents Triton X-100, Tween 20, Brij-35, EDTA, N -Cetyl-N ,N ,N -trimethylammonium bromide (CTAB), ortho-phenylenediamine (OPD), Coomassie Blue R-250, hyaluronic acid, diaminobenzidine (DAB), phenylmethanesulfonyl fluoride (PMSF), 1,10-phenanthroline, bovine serum albumin (BSA), fibrinogen from human plasma, gelatin type A, goat anti-horse (GAH) IgG horseradish peroxidase (IgGHRPO), concanavalin A (Con A), and Wheat Germ Agglutinin (WGA) labelled with peroxidase, were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. Dithiothreitol (DTT) and iodoacetamide were obtained from GE Healthcare Life Sciences (Pittsburgh, PA, USA).

    Diafiltration Assay:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: SDS-PAGE and Western Blot analyzed fractions were pooled, buffer exchanged and dialyzed using Vivaspin centrifugal units coupled to diafiltration cups (Sartorius, #Cat. VSA005) against acetate buffer (50 mM, pH 5). .. Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    MTT Assay:

    Article Title: Enzymatic and Pro-Inflammatory Activities of Bothrops lanceolatus Venom: Relevance for Envenomation
    Article Snippet: Chemicals and Reagents Triton X-100, Tween 20, Brij-35, EDTA, N -Cetyl-N ,N ,N -trimethylammonium bromide (CTAB), ortho-phenylenediamine (OPD), Coomassie Blue R-250, hyaluronic acid, diaminobenzidine (DAB), phenylmethanesulfonyl fluoride (PMSF), 1,10-phenanthroline, bovine serum albumin (BSA), fibrinogen from human plasma, gelatin type A, goat anti-horse (GAH) IgG horseradish peroxidase (IgGHRPO), concanavalin A (Con A), and Wheat Germ Agglutinin (WGA) labelled with peroxidase, were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. 3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were from Merck (Darmstadt, Germany).

    BIA-KA:

    Article Title: Heterologous Expression and Purification of Active Divercin V41, a Class IIa Bacteriocin Encoded by a Synthetic Gene in Escherichia coli
    Article Snippet: Proteins in glycine-SDS-PAGE gels were stained with Coomassie blue R-250 (Sigma). .. The protein concentration was determined by using the BCA protein assay reagent (Pierce, Rockford, Ill.), with bovine serum albumin as a standard.

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: Total protein determination was achieved via the Pierce BCA assay. .. The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein.

    Immunostaining:

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: Experimental Procedures Methods for construction of recombinant adenovirus vectors containing CMV promoter-driven ATP7A or ATP7B cDNA fused with a 3′ c-myc tag (rAdATP7Amyc or rAdATP7Bmyc), infection of COS-1 cells, and immunostaining for detection of the heterologous ATP7A or ATP7B protein, were previously described in detail for ATP7B ( ). .. The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein.

    Electrophoresis:

    Article Title: Diverse matrix metalloproteinase functions regulate cancer amoeboid migration
    Article Snippet: Following electrophoresis, SDS was washed out by incubating the gels in 2.5% Triton X-100 for 30 min at room temperature. .. Gels were then washed three times in distilled water and incubated in substrate buffer (50 mM (Tris pH 7.4), 200 mM NaCl, 2 mM CaCl2 , 1 mM MgCl2 ) at room temperature for 30 min and then at 37 °C for 16 h. Gels were stained with 0.5% Coomassie Blue R-250 (Sigma) and destained in 5% methanol and 10% acetic acid.

    Article Title: Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry
    Article Snippet: Following electrophoresis, the gels were fixed for 1 h in a solution of 40% (v/v) aqueous ethanol (99.8%; Sigma-Aldrich) and 10% (v/v) acetic acid (Merck Millipore, Darmstadt, Germany) at room temperature. .. The gels were then washed for 30 min in fresh fixing solution and incubated with Coomassie Blue R-250 0.2% diluted in fixative solution for 2 h at room temperature (Sigma-Aldrich).

    Article Title: Ovulation-inducing factor: a protein component of llama seminal plasma
    Article Snippet: To examine the effects of treatment on seminal plasma protein band profiles, samples from each treatment were reduced, denatured, and separated by electrophoresis on 12% polyacrylamide gel (SDS-PAGE) based on the protocol of Laemmli [ ]. .. Gels were stained with Coomassie Blue R-250 (Sigma-Aldrich, St Louis, Missouri, USA).

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma). .. We estimated the yield of proteins by comparing quantities of proteins extracted using our protocol and a direct boiling in 2.5% SDS-containing electrophoresis sample buffer, followed by 1D SDS-PAGE.

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: After electrophoresis, the gels were washed with 2.5% Triton X-100 for 30 min at room temperature and then incubated overnight at 37°C in 100 m m Tris buffer, pH 7.4, containing 15 m m CaCl2 . .. They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid.

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: SDS gel electrophoresis was performed by the method of Laemmli ( ) at pH 8.3 or Weber and Osborn ( ) at pH 6.3. .. The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein.

    Article Title: Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi
    Article Snippet: For gel electrophoresis, proteins were heated to 85°C for 5 min in Novex 2× Tricine-sodium dodecyl sulfate (SDS) sample buffer (Invitrogen, Carlsbad, Calif.) or to 70°C for 10 min in Novex 4× NuPAGE sample buffer and separated through Tricine-10 to 20% polyacrylamide gradient gels or through 4 to 12% NuPAGE bis-Tris gels by using a Novex XCell Sure Lock electrophoresis cell (Invitrogen). .. Proteins were stained with Coomassie blue R-250 (Sigma).

    Incubation:

    Article Title: Diverse matrix metalloproteinase functions regulate cancer amoeboid migration
    Article Snippet: .. Gels were then washed three times in distilled water and incubated in substrate buffer (50 mM (Tris pH 7.4), 200 mM NaCl, 2 mM CaCl2 , 1 mM MgCl2 ) at room temperature for 30 min and then at 37 °C for 16 h. Gels were stained with 0.5% Coomassie Blue R-250 (Sigma) and destained in 5% methanol and 10% acetic acid. ..

    Article Title: Protein A-Mouse Acidic Mammalian Chitinase-V5-His Expressed in Periplasmic Space of Escherichia coli Possesses Chitinase Functions Comparable to CHO-Expressed Protein
    Article Snippet: The proteins in the gel were visualized by staining with Coomassie Blue R-250 (Sigma-Aldrich). .. Separated proteins were electrophoretically transferred to a PVDF (polyvinylidene fluoride) membrane (Immobilon-P, Millipore), which was incubated with an anti-V5-HRP monoclonal antibody (Invitrogen).

    Article Title: Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry
    Article Snippet: .. The gels were then washed for 30 min in fresh fixing solution and incubated with Coomassie Blue R-250 0.2% diluted in fixative solution for 2 h at room temperature (Sigma-Aldrich). .. The gels were destained using fixative solution for 2 h, followed by incubation in water at room temperature until complete destaining.

    Article Title: Ovulation-inducing factor: a protein component of llama seminal plasma
    Article Snippet: For the control group, seminal plasma was incubated in a shaker water bath at 38°C for 1 hour without the presence of proteinase-K, and PMSF was added to a final concentration of 5 mM. .. Gels were stained with Coomassie Blue R-250 (Sigma-Aldrich, St Louis, Missouri, USA).

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: After electrophoresis, the gels were washed with 2.5% Triton X-100 for 30 min at room temperature and then incubated overnight at 37°C in 100 m m Tris buffer, pH 7.4, containing 15 m m CaCl2 . .. They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid.

    Activity Assay:

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: One hundred fifty milligrams of tissue was mechanically dispersed, placed in a culture dish without rinsing, and incubated for 12 hr with 500 μl of DMEM medium (Life Technologies, Paisley, UK) without serum; this step allowed us to detect the activity of secreted MMPs 2 and 9 in the conditioned medium. .. They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid.

    Expressing:

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: The sections of the biopsies have to be stored for a validation study, when expression of proteins of interest identified by proteomics would be monitored by immunohistochemistry with specific antibodies in the sections of the same sample. .. The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma).

    Bradford Assay:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: Total protein concentrations of crude supernatant or purified fractions were determined by Bradford assay [ ] using Bovine Serum Albumin (BSA) as calibration standard. .. Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    Zymography:

    Article Title: Diverse matrix metalloproteinase functions regulate cancer amoeboid migration
    Article Snippet: Paragraph title: Gelatin zymography ... Gels were then washed three times in distilled water and incubated in substrate buffer (50 mM (Tris pH 7.4), 200 mM NaCl, 2 mM CaCl2 , 1 mM MgCl2 ) at room temperature for 30 min and then at 37 °C for 16 h. Gels were stained with 0.5% Coomassie Blue R-250 (Sigma) and destained in 5% methanol and 10% acetic acid.

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: Paragraph title: Gelatin zymography ... They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid.

    Article Title: Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors
    Article Snippet: Paragraph title: 2.1 Materials for Zymography Experiment ... Staining solution: 0.5% w/v or 2 g Coomassie blue R-250 (Sigma), 25% v/v or 100 ml isopropanol (Sigma), and 10% v/v or 40 ml acetic acid (Fisher Scientific) to 260 ml dH2 O. Filter through filter paper before use.

    High Performance Liquid Chromatography:

    Article Title: Heterologous Expression and Purification of Active Divercin V41, a Class IIa Bacteriocin Encoded by a Synthetic Gene in Escherichia coli
    Article Snippet: Proteins in glycine-SDS-PAGE gels were stained with Coomassie blue R-250 (Sigma). .. The purity of DvnV41 and DvnRV41 was analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC) using a C18 nucleosyl column (250 by 4.6 nm, 5-μm-diameter particles, column 300A; CIL, Sainte Foy la Grande, France).

    Immunohistochemistry:

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: The sections of the biopsies have to be stored for a validation study, when expression of proteins of interest identified by proteomics would be monitored by immunohistochemistry with specific antibodies in the sections of the same sample. .. The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma).

    Concentration Assay:

    Article Title: Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry
    Article Snippet: Each sample (20 μ g of each) was mixed with the sample buffer to a final concentration of 0.06 M Tris-HCl pH 6.8, 2% SDS, 5% β-mercaptoethanol, 10% glycerol and 0.025% bromophenol blue. .. The gels were then washed for 30 min in fresh fixing solution and incubated with Coomassie Blue R-250 0.2% diluted in fixative solution for 2 h at room temperature (Sigma-Aldrich).

    Article Title: Ovulation-inducing factor: a protein component of llama seminal plasma
    Article Snippet: For the control group, seminal plasma was incubated in a shaker water bath at 38°C for 1 hour without the presence of proteinase-K, and PMSF was added to a final concentration of 5 mM. .. Gels were stained with Coomassie Blue R-250 (Sigma-Aldrich, St Louis, Missouri, USA).

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: We prepared experiments with buffers that contained different detergents and concentration , because different detergents solubilize cells membranes with different efficiencies., Extraction conditions (time, treatments) were similar for all tested conditions. .. The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma).

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein. .. Tergitol-type NP-40 was then added to a final concentration of 1%, followed by 2500 units of endoglycosidase PNGase F (NEB).

    Article Title: Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors
    Article Snippet: Staining solution: 0.5% w/v or 2 g Coomassie blue R-250 (Sigma), 25% v/v or 100 ml isopropanol (Sigma), and 10% v/v or 40 ml acetic acid (Fisher Scientific) to 260 ml dH2 O. Filter through filter paper before use. .. Final concentration 50% v/v methanol (Sigma) and 10% v/v acetic acid in dH2 O. Homogenization buffer: 0.02 M or 0.418 g 3-[N-Morpholino]propanesulfonicacid (MOPS) (Sigma), 4% w/v or 4 g SDS, 10% v/v or 10 ml glycerol, and 90 ml dH2 O.

    Protease Inhibitor:

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein. .. In some experiments, before electrophoresis, we subjected the microsomal protein to deglycosylation by dissolving 50 μg of microsomal protein in 40 μL of reaction buffer containing 50 mM sodium phosphate, 0.25% SDS, 20 mM DTT, and protease inhibitor cocktail (Sigma) (pH 7.5).

    Infection:

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: Preparation of the microsomal fraction from infected COS-1 cells was also described previously ( ). .. The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein.

    Inhibition:

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid. .. For inhibition of metalloproteinase activity, the gels were incubated with 4 m m 1,10-pheanthroline as described previously ( ).

    Protein Concentration:

    Article Title: Heterologous Expression and Purification of Active Divercin V41, a Class IIa Bacteriocin Encoded by a Synthetic Gene in Escherichia coli
    Article Snippet: Proteins in glycine-SDS-PAGE gels were stained with Coomassie blue R-250 (Sigma). .. The protein concentration was determined by using the BCA protein assay reagent (Pierce, Rockford, Ill.), with bovine serum albumin as a standard.

    Article Title: Protein A-Mouse Acidic Mammalian Chitinase-V5-His Expressed in Periplasmic Space of Escherichia coli Possesses Chitinase Functions Comparable to CHO-Expressed Protein
    Article Snippet: Paragraph title: Determination of Protein Concentration, SDS-polyacrylamide Gel Electrophoresis and Western Blotting ... The proteins in the gel were visualized by staining with Coomassie Blue R-250 (Sigma-Aldrich).

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: Briefly, equal volumes (20 μl) of the conditioned medium, normalized for protein concentration, were mixed 1:4 with 4× SDS sample buffer, and 25 μl of the mixture was loaded into a well of a 10% zymography gel copolymerized with 0.1% gelatin. .. They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid.

    Article Title: Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors
    Article Snippet: When ready to load the samples in the gel and depending on the protein concentration in the sample tissue homogenate, add one part of the sample to one part sample buffer. .. Staining solution: 0.5% w/v or 2 g Coomassie blue R-250 (Sigma), 25% v/v or 100 ml isopropanol (Sigma), and 10% v/v or 40 ml acetic acid (Fisher Scientific) to 260 ml dH2 O. Filter through filter paper before use.

    Recombinant:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: Molecular mass estimation of recombinant ScExlx1 was done loading 5 μg of protein into 12% SDS-polyacrilamyde gel. .. Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: Experimental Procedures Methods for construction of recombinant adenovirus vectors containing CMV promoter-driven ATP7A or ATP7B cDNA fused with a 3′ c-myc tag (rAdATP7Amyc or rAdATP7Bmyc), infection of COS-1 cells, and immunostaining for detection of the heterologous ATP7A or ATP7B protein, were previously described in detail for ATP7B ( ). .. The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein.

    Molecular Weight:

    Article Title: Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry
    Article Snippet: The molecular weight standard was BenchMark Protein Ladder (Invitrogen Life Technologies, Carlsbad, CA, USA). .. The gels were then washed for 30 min in fresh fixing solution and incubated with Coomassie Blue R-250 0.2% diluted in fixative solution for 2 h at room temperature (Sigma-Aldrich).

    Nucleic Acid Electrophoresis:

    Article Title: Heterologous Expression and Purification of Active Divercin V41, a Class IIa Bacteriocin Encoded by a Synthetic Gene in Escherichia coli
    Article Snippet: Proteins were separated under reducing conditions by Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE) (16.5% polyacrylamide [Sigma]) ( ) or glycine-SDS-PAGE (15% polyacrylamide) ( ). .. Proteins in glycine-SDS-PAGE gels were stained with Coomassie blue R-250 (Sigma).

    Article Title: Purification and Molecular Characterization of a Tripeptidase (PepT) from Lactobacillus helveticus
    Article Snippet: After each purification step, the purity of PepT active fractions was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as described by Laemmli , with gels containing 12% acrylamide. .. Gels were stained with Coomassie blue R 250 (Sigma), and the molecular mass of the denaturated PepT was estimated by using a low-molecular-weight protein standard (Pharmacia Biotech).

    Article Title: Protein A-Mouse Acidic Mammalian Chitinase-V5-His Expressed in Periplasmic Space of Escherichia coli Possesses Chitinase Functions Comparable to CHO-Expressed Protein
    Article Snippet: Paragraph title: Determination of Protein Concentration, SDS-polyacrylamide Gel Electrophoresis and Western Blotting ... The proteins in the gel were visualized by staining with Coomassie Blue R-250 (Sigma-Aldrich).

    Article Title: Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi
    Article Snippet: For gel electrophoresis, proteins were heated to 85°C for 5 min in Novex 2× Tricine-sodium dodecyl sulfate (SDS) sample buffer (Invitrogen, Carlsbad, Calif.) or to 70°C for 10 min in Novex 4× NuPAGE sample buffer and separated through Tricine-10 to 20% polyacrylamide gradient gels or through 4 to 12% NuPAGE bis-Tris gels by using a Novex XCell Sure Lock electrophoresis cell (Invitrogen). .. Proteins were stained with Coomassie blue R-250 (Sigma).

    Western Blot:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: Paragraph title: Protein purification and quantification. SDS-PAGE and Western Blot ... Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    Article Title: Protein A-Mouse Acidic Mammalian Chitinase-V5-His Expressed in Periplasmic Space of Escherichia coli Possesses Chitinase Functions Comparable to CHO-Expressed Protein
    Article Snippet: Paragraph title: Determination of Protein Concentration, SDS-polyacrylamide Gel Electrophoresis and Western Blotting ... The proteins in the gel were visualized by staining with Coomassie Blue R-250 (Sigma-Aldrich).

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein. .. In addition, the protein was transferred from the gels to a PDVF membrane for Western blotting, which was then visualized using 9E10 monoclonal antibodies against the c-myc tag.

    Nickel Column:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: The concentrated supernatant was loaded on a HisTrap excel Nickel column (GE Healtcare, #Cat. 17-3712-05) connected to a peristaltic pump and previously equilibrated with phosphate buffer (20 mM NaH2 PO4 , 20 mM Imidazole, 0.5 M NaCl pH 7.4). .. Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    Filtration:

    Article Title: Purification and Molecular Characterization of a Tripeptidase (PepT) from Lactobacillus helveticus
    Article Snippet: Gels were stained with Coomassie blue R 250 (Sigma), and the molecular mass of the denaturated PepT was estimated by using a low-molecular-weight protein standard (Pharmacia Biotech). .. The relative molecular mass of the purified PepT (10 μg) in native form was determined by gel filtration on a Superdex 200 HR10/30 column, using the running conditions described above for gel filtration on Superdex 200.

    Purification:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: Total protein concentrations of crude supernatant or purified fractions were determined by Bradford assay [ ] using Bovine Serum Albumin (BSA) as calibration standard. .. Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    Article Title: Purification and Molecular Characterization of a Tripeptidase (PepT) from Lactobacillus helveticus
    Article Snippet: After each purification step, the purity of PepT active fractions was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as described by Laemmli , with gels containing 12% acrylamide. .. Gels were stained with Coomassie blue R 250 (Sigma), and the molecular mass of the denaturated PepT was estimated by using a low-molecular-weight protein standard (Pharmacia Biotech).

    Protein Purification:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: Paragraph title: Protein purification and quantification. SDS-PAGE and Western Blot ... Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    Immunodetection:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation. .. The membrane was blocked with PBST plus skimmed milk (3%) for 20 min and washed with PBST. c-Myc (9E10) (Santa Cruz Biotechnology, #Cat. Sc-40) and anti-His antibodies (Roche, #Cat. 11922416001) were used for immunodetection (dilution 1:5000) and signal detection was visualized using an anti-mouse alkaline phosphatase conjugate (Sigma, #Cat. A3562) (dilution 1:10000) by incubating 30 min in PBST plus skimmed milk.

    Fast Protein Liquid Chromatography:

    Article Title: Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi
    Article Snippet: Proteins in peak FPLC fractions were concentrated by precipitation with a 2.5-fold excess of ice-cold ethanol. .. Proteins were stained with Coomassie blue R-250 (Sigma).

    Polyacrylamide Gel Electrophoresis:

    Article Title: Protein A-Mouse Acidic Mammalian Chitinase-V5-His Expressed in Periplasmic Space of Escherichia coli Possesses Chitinase Functions Comparable to CHO-Expressed Protein
    Article Snippet: The protein fractions that were obtained as described above were analyzed using standard SDS-polyacrylamide gel electrophoresis (PAGE) . .. The proteins in the gel were visualized by staining with Coomassie Blue R-250 (Sigma-Aldrich).

    Staining:

    Article Title: Identification of Altered Plasma Proteins by Proteomic Study in Valvular Heart Diseases and the Potential Clinical Significance
    Article Snippet: .. Image Analysis After 2-DE separation, the nine gels were stained with Coomassie blue R-250 (Sigma, St Louis, MO, USA). .. Spot detection and quantification were carried out using PDQuest 2D-analysis software (Bio-Rad, Hercules, CA).

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: .. Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation. .. For Western blot analysis, purified ScExlx1 was run on a 12% SDS-PAGE and blotted onto a nitrocellulose membrane (Bio-Rad) using a wet tank blotting system (Bio-Rad).

    Article Title: Heterologous Expression and Purification of Active Divercin V41, a Class IIa Bacteriocin Encoded by a Synthetic Gene in Escherichia coli
    Article Snippet: .. Proteins in glycine-SDS-PAGE gels were stained with Coomassie blue R-250 (Sigma). .. An ultra-low-range marker (Sigma) was used as a molecular mass marker (26.6, 17.0, 14.2, 6.5, 3.5, and 1.1 kDa).

    Article Title: Diverse matrix metalloproteinase functions regulate cancer amoeboid migration
    Article Snippet: .. Gels were then washed three times in distilled water and incubated in substrate buffer (50 mM (Tris pH 7.4), 200 mM NaCl, 2 mM CaCl2 , 1 mM MgCl2 ) at room temperature for 30 min and then at 37 °C for 16 h. Gels were stained with 0.5% Coomassie Blue R-250 (Sigma) and destained in 5% methanol and 10% acetic acid. ..

    Article Title: Purification and Molecular Characterization of a Tripeptidase (PepT) from Lactobacillus helveticus
    Article Snippet: .. Gels were stained with Coomassie blue R 250 (Sigma), and the molecular mass of the denaturated PepT was estimated by using a low-molecular-weight protein standard (Pharmacia Biotech). .. The relative molecular mass of the purified PepT (10 μg) in native form was determined by gel filtration on a Superdex 200 HR10/30 column, using the running conditions described above for gel filtration on Superdex 200.

    Article Title: Protein A-Mouse Acidic Mammalian Chitinase-V5-His Expressed in Periplasmic Space of Escherichia coli Possesses Chitinase Functions Comparable to CHO-Expressed Protein
    Article Snippet: .. The proteins in the gel were visualized by staining with Coomassie Blue R-250 (Sigma-Aldrich). .. Separated proteins were electrophoretically transferred to a PVDF (polyvinylidene fluoride) membrane (Immobilon-P, Millipore), which was incubated with an anti-V5-HRP monoclonal antibody (Invitrogen).

    Article Title: Ovulation-inducing factor: a protein component of llama seminal plasma
    Article Snippet: .. Gels were stained with Coomassie Blue R-250 (Sigma-Aldrich, St Louis, Missouri, USA). .. Experiment 3 - treatment with pronase E The experiment was conducted from May to June at the University of Saskatchewan, Canada.

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: .. The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma). .. To evaluate the maximal level of protein extraction, a similar sample was extracted using SDS.

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: .. They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid. .. Enzyme activity, attributable to MMPs 2 and 9 on the basis of their respective molecular weights, was visualized in the gelatin-containing zymograms as clear bands on a blue background.

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: .. The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein. .. In addition, the protein was transferred from the gels to a PDVF membrane for Western blotting, which was then visualized using 9E10 monoclonal antibodies against the c-myc tag.

    Article Title: Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors
    Article Snippet: .. Staining solution: 0.5% w/v or 2 g Coomassie blue R-250 (Sigma), 25% v/v or 100 ml isopropanol (Sigma), and 10% v/v or 40 ml acetic acid (Fisher Scientific) to 260 ml dH2 O. Filter through filter paper before use. ..

    Article Title: Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi
    Article Snippet: .. Proteins were stained with Coomassie blue R-250 (Sigma). .. For immunoblotting, proteins were transferred to a polyvinylidene difluoride membrane (PALL Corporation, Ann Arbor, Mich.) and probed with antibodies.

    SDS Page:

    Article Title: A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune
    Article Snippet: Paragraph title: Protein purification and quantification. SDS-PAGE and Western Blot ... Protein bands were visualized by Coomassie Blue R-250 (Sigma-Aldrich) staining and PageRuler Plus Pre-stained Protein Ladder (Thermo Scientific, #Cat. 26619) was used for molecular mass estimation.

    Article Title: Diverse matrix metalloproteinase functions regulate cancer amoeboid migration
    Article Snippet: Secreted serum-free medium from cells grown on plastic for 48 h was loaded on 10% SDS-PAGE gels containing 1 mg ml−1 gelatin (Sigma) in non-reduced protein sample buffer. .. Gels were then washed three times in distilled water and incubated in substrate buffer (50 mM (Tris pH 7.4), 200 mM NaCl, 2 mM CaCl2 , 1 mM MgCl2 ) at room temperature for 30 min and then at 37 °C for 16 h. Gels were stained with 0.5% Coomassie Blue R-250 (Sigma) and destained in 5% methanol and 10% acetic acid.

    Article Title: Purification and Molecular Characterization of a Tripeptidase (PepT) from Lactobacillus helveticus
    Article Snippet: After each purification step, the purity of PepT active fractions was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as described by Laemmli , with gels containing 12% acrylamide. .. Gels were stained with Coomassie blue R 250 (Sigma), and the molecular mass of the denaturated PepT was estimated by using a low-molecular-weight protein standard (Pharmacia Biotech).

    Article Title: Ovulation-inducing factor: a protein component of llama seminal plasma
    Article Snippet: To examine the effects of treatment on seminal plasma protein band profiles, samples from each treatment were reduced, denatured, and separated by electrophoresis on 12% polyacrylamide gel (SDS-PAGE) based on the protocol of Laemmli [ ]. .. Gels were stained with Coomassie Blue R-250 (Sigma-Aldrich, St Louis, Missouri, USA).

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: .. The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma). .. To evaluate the maximal level of protein extraction, a similar sample was extracted using SDS.

    Software:

    Article Title: Identification of Altered Plasma Proteins by Proteomic Study in Valvular Heart Diseases and the Potential Clinical Significance
    Article Snippet: Image Analysis After 2-DE separation, the nine gels were stained with Coomassie blue R-250 (Sigma, St Louis, MO, USA). .. Spot detection and quantification were carried out using PDQuest 2D-analysis software (Bio-Rad, Hercules, CA).

    SDS-Gel:

    Article Title: Comparative Features of Copper ATPases ATP7A and ATP7B Heterologously Expressed in COS-1 Cells
    Article Snippet: SDS gel electrophoresis was performed by the method of Laemmli ( ) at pH 8.3 or Weber and Osborn ( ) at pH 6.3. .. The gels were stained with either Coomassie Blue R-250 (Sigma) for detection of protein bands or Pro-Q Emerald (Invitrogen) for detection of glycoprotein.

    Selection:

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: The first step in protein extraction from the tissue is selection of an optimal extraction buffer. .. The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma).

    Homogenization:

    Article Title: Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors
    Article Snippet: Staining solution: 0.5% w/v or 2 g Coomassie blue R-250 (Sigma), 25% v/v or 100 ml isopropanol (Sigma), and 10% v/v or 40 ml acetic acid (Fisher Scientific) to 260 ml dH2 O. Filter through filter paper before use. .. Final concentration 50% v/v methanol (Sigma) and 10% v/v acetic acid in dH2 O. Homogenization buffer: 0.02 M or 0.418 g 3-[N-Morpholino]propanesulfonicacid (MOPS) (Sigma), 4% w/v or 4 g SDS, 10% v/v or 10 ml glycerol, and 90 ml dH2 O.

    Protein Extraction:

    Article Title: Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry
    Article Snippet: Paragraph title: Protein extraction and gels ... The gels were then washed for 30 min in fresh fixing solution and incubated with Coomassie Blue R-250 0.2% diluted in fixative solution for 2 h at room temperature (Sigma-Aldrich).

    Article Title: Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis
    Article Snippet: The first step in protein extraction from the tissue is selection of an optimal extraction buffer. .. The output of experiments was monitored by intensity of prepared and separated in 1D SDS-PAGE gels proteins, stained with coomassie blue R-250 (Sigma).

    Protein Electrophoresis:

    Article Title: Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi
    Article Snippet: Paragraph title: Protein electrophoresis, immunoblotting, and antiserum. ... Proteins were stained with Coomassie blue R-250 (Sigma).

    Marker:

    Article Title: Heterologous Expression and Purification of Active Divercin V41, a Class IIa Bacteriocin Encoded by a Synthetic Gene in Escherichia coli
    Article Snippet: Proteins in glycine-SDS-PAGE gels were stained with Coomassie blue R-250 (Sigma). .. An ultra-low-range marker (Sigma) was used as a molecular mass marker (26.6, 17.0, 14.2, 6.5, 3.5, and 1.1 kDa).

    Article Title: Diverse matrix metalloproteinase functions regulate cancer amoeboid migration
    Article Snippet: Gels were then washed three times in distilled water and incubated in substrate buffer (50 mM (Tris pH 7.4), 200 mM NaCl, 2 mM CaCl2 , 1 mM MgCl2 ) at room temperature for 30 min and then at 37 °C for 16 h. Gels were stained with 0.5% Coomassie Blue R-250 (Sigma) and destained in 5% methanol and 10% acetic acid. .. In melanoma cells, MMP-9 82-kDa form was detected just right above the 76-kDa marker, while MMP-2 62-kDa form was detected above the 52-kDa marker.

    Lysis:

    Article Title: Spatiotemporal Expression Patterns of Metalloproteinases and Their Inhibitors in the Postnatal Developing Rat Cerebellum
    Article Snippet: Cell loss by lysis was quantified using Trypan blue and was minimal after tissue treatment ( < 10%). .. They were then fixed with 30% methanol–10% acetic acid, stained with 0.25% Coomassie blue R-250 (Sigma, St. Louis, MO), and destained with 10% methanol–10% acetic acid.

    other:

    Article Title: Contribution of Amino Acid Region 334-335 from Factor Va Heavy Chain to the Catalytic Efficiency of Prothrombinase †
    Article Snippet: Materials, Reagents, and Proteins Diisopropyl fluorophosphate (DFP), O -phenylenediamine (OPD) dihydrochloride, N -(2-hydroxyethyl)piperazine-N ′-2-ethanesulfonic acid (Hepes), Trizma (Tris base), and Coomassie Blue R-250 were purchased from Sigma (St. Louis, MO).

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    Millipore coomassie brilliant blue r 250
    A high molecular weight complex (HX) showing Pho1 activity was found in the sweet potato roots. ( A ) The protein fraction from the extract of sweet potato roots was resolved by size-exclusion chromatography (Sephacryl S-300). The fractions (nos. 33∼71) after chromatography were analyzed by 6% native PAGE, and then stained with <t>Coomassie</t> Brilliant Blue R-250 (Coomassie). The molecular mass markers (thyroglobulin (porcine thyroid ), 669 kDa; ferritin (equine spleen ), 440 kDa; catalase (bovine liver ), 232 kDa) were revealed in lane 1. ( B ) The native PAGE gel was incubated with the reaction buffer containing 20 mM MES-KOH (pH 5.9), 1.2% soluble starch and 32 mM Glc-1-P at 37°C for 2 h. The zymogram was detected using iodine staining to reveal the catalytic activity of Pho1 in the direction of starch synthesis. Both of Pho1 and HX showed starch-synthesizing activity. The bands with negative staining were identified as beta amylase (BA). ( C ) The native PAGE gel was transferred to PVDF membranes and subsequently analyzed by western boltting with anti-20S proteasome antibody (α20S). The molecular mass in kilodalton (kDa) was shown as indicated on the bottom of the figure by applying the calibration of the size-exclusion chromatography column with known molecular mass standards (thyroglobulin (bovine), 670 kDa; γ-globulin (bovine), 158 kDa; ovalbumin (chicken), 44 kDa).
    Coomassie Brilliant Blue R 250, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 376 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/coomassie brilliant blue r 250/product/Millipore
    Average 99 stars, based on 376 article reviews
    Price from $9.99 to $1999.99
    coomassie brilliant blue r 250 - by Bioz Stars, 2020-04
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    92
    Millipore coomassie blue r 250 staining
    Fig. 1. TbSP1 identification. ( A ) Equal amounts of SLM-released protein were subjected to SDS–PAGE and stained with <t>Coomassie</t> Blue R-250. Days of in vitro culture (d) and the migration positions of molecular mass markers are indicated at the top and at the left, respectively. The migration position (p23) and the N-terminal sequence (p23/20) of the TbSP1 polypeptide are shown on the right. ( B ) Genomic DNA digested with Eco RI (lane 1), Bam HI (lane 2) or Hin dIII (lane 3) was probed with the TbSP1 cDNA. The migration positions of DNA size markers are indicated on the left. ( C ) Balanced amounts of total RNA extracted from synthetic solid (SSM) or liquid (SLM) mycelial cultures were gel fractionated and probed with the 32 P-labeled TbSP1 cDNA. The migration positions and the amounts of the 28S and 18S rRNAs, utilized as internal references, are shown on the right and in the lower panel, respectively.
    Coomassie Blue R 250 Staining, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/coomassie blue r 250 staining/product/Millipore
    Average 92 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    A high molecular weight complex (HX) showing Pho1 activity was found in the sweet potato roots. ( A ) The protein fraction from the extract of sweet potato roots was resolved by size-exclusion chromatography (Sephacryl S-300). The fractions (nos. 33∼71) after chromatography were analyzed by 6% native PAGE, and then stained with Coomassie Brilliant Blue R-250 (Coomassie). The molecular mass markers (thyroglobulin (porcine thyroid ), 669 kDa; ferritin (equine spleen ), 440 kDa; catalase (bovine liver ), 232 kDa) were revealed in lane 1. ( B ) The native PAGE gel was incubated with the reaction buffer containing 20 mM MES-KOH (pH 5.9), 1.2% soluble starch and 32 mM Glc-1-P at 37°C for 2 h. The zymogram was detected using iodine staining to reveal the catalytic activity of Pho1 in the direction of starch synthesis. Both of Pho1 and HX showed starch-synthesizing activity. The bands with negative staining were identified as beta amylase (BA). ( C ) The native PAGE gel was transferred to PVDF membranes and subsequently analyzed by western boltting with anti-20S proteasome antibody (α20S). The molecular mass in kilodalton (kDa) was shown as indicated on the bottom of the figure by applying the calibration of the size-exclusion chromatography column with known molecular mass standards (thyroglobulin (bovine), 670 kDa; γ-globulin (bovine), 158 kDa; ovalbumin (chicken), 44 kDa).

    Journal: PLoS ONE

    Article Title: Plastidial Starch Phosphorylase in Sweet Potato Roots Is Proteolytically Modified by Protein-Protein Interaction with the 20S Proteasome

    doi: 10.1371/journal.pone.0035336

    Figure Lengend Snippet: A high molecular weight complex (HX) showing Pho1 activity was found in the sweet potato roots. ( A ) The protein fraction from the extract of sweet potato roots was resolved by size-exclusion chromatography (Sephacryl S-300). The fractions (nos. 33∼71) after chromatography were analyzed by 6% native PAGE, and then stained with Coomassie Brilliant Blue R-250 (Coomassie). The molecular mass markers (thyroglobulin (porcine thyroid ), 669 kDa; ferritin (equine spleen ), 440 kDa; catalase (bovine liver ), 232 kDa) were revealed in lane 1. ( B ) The native PAGE gel was incubated with the reaction buffer containing 20 mM MES-KOH (pH 5.9), 1.2% soluble starch and 32 mM Glc-1-P at 37°C for 2 h. The zymogram was detected using iodine staining to reveal the catalytic activity of Pho1 in the direction of starch synthesis. Both of Pho1 and HX showed starch-synthesizing activity. The bands with negative staining were identified as beta amylase (BA). ( C ) The native PAGE gel was transferred to PVDF membranes and subsequently analyzed by western boltting with anti-20S proteasome antibody (α20S). The molecular mass in kilodalton (kDa) was shown as indicated on the bottom of the figure by applying the calibration of the size-exclusion chromatography column with known molecular mass standards (thyroglobulin (bovine), 670 kDa; γ-globulin (bovine), 158 kDa; ovalbumin (chicken), 44 kDa).

    Article Snippet: After electrophoresis, proteins were either stained with Coomassie Brilliant Blue R-250 or transferred to the PVDF membrane (Immobilon-P, Millipore) for western blotting.

    Techniques: Molecular Weight, Activity Assay, Size-exclusion Chromatography, Chromatography, Clear Native PAGE, Staining, Incubation, Gas Chromatography, Negative Staining, Western Blot

    Dpe1 was co-immunoprecipitated with Pho1 by using anti-Pho1 antibody. (A) Whole cell extracts from the 10-week sweet potato storage roots (protein concentration 1 mg/ml) were pre-incubated with glucan-degrading enzymes (5 U/mL of amyloglucosidase and 7.7 U/mL of α-amylase) at 25°C for 20 min, and then incubated with antibodies for Protein-A Sepharose immunoprecipitation: lane 1, conventional antisera against Pho1 (αPho1); lane 2, Pho1 specific mAb (H7c); lane 3, an unrelated mAb (Mock); lane 4, the control experiment using Protein A-Sepharose beads alone without any antibody (ProA). After the immunoprecipitation procedures, the eluted proteins were directly analyzed on 12.5% SDS-PAGE, and stained with Coomassie Brilliant Blue R-250 (CBR). For comparison, 10 μg of the whole cell extracts were also analyzed on the same SDS-PAGE (designated as Input). Arrowheads indicated the positions of Pho1 (closed) and Dpe1 (open). M, pre-stained SDS-PAGE molecular mass standards. (B) The protein samples from bands a and b were sliced out for mass spectrometry analysis. The results demonstrated that band a is Pho1 and band b is Dpe1.

    Journal: PLoS ONE

    Article Title: Plastidial α-glucan phosphorylase 1 complexes with disproportionating enzyme 1 in Ipomoea batatas storage roots for elevating malto-oligosaccharide metabolism

    doi: 10.1371/journal.pone.0177115

    Figure Lengend Snippet: Dpe1 was co-immunoprecipitated with Pho1 by using anti-Pho1 antibody. (A) Whole cell extracts from the 10-week sweet potato storage roots (protein concentration 1 mg/ml) were pre-incubated with glucan-degrading enzymes (5 U/mL of amyloglucosidase and 7.7 U/mL of α-amylase) at 25°C for 20 min, and then incubated with antibodies for Protein-A Sepharose immunoprecipitation: lane 1, conventional antisera against Pho1 (αPho1); lane 2, Pho1 specific mAb (H7c); lane 3, an unrelated mAb (Mock); lane 4, the control experiment using Protein A-Sepharose beads alone without any antibody (ProA). After the immunoprecipitation procedures, the eluted proteins were directly analyzed on 12.5% SDS-PAGE, and stained with Coomassie Brilliant Blue R-250 (CBR). For comparison, 10 μg of the whole cell extracts were also analyzed on the same SDS-PAGE (designated as Input). Arrowheads indicated the positions of Pho1 (closed) and Dpe1 (open). M, pre-stained SDS-PAGE molecular mass standards. (B) The protein samples from bands a and b were sliced out for mass spectrometry analysis. The results demonstrated that band a is Pho1 and band b is Dpe1.

    Article Snippet: After electrophoresis, proteins were either stained with Coomassie Brilliant Blue R-250 (CBR) or transferred to the PVDF membranes (Immobilon-P, Millipore) for western blotting.

    Techniques: Immunoprecipitation, Protein Concentration, Incubation, SDS Page, Staining, Mass Spectrometry

    SDS–PAGE analysis of released mouse ST6Gal-I with or without β-mercaptoethanol (β-ME) treatment. Twenty microgram of released mouse ST6Gal-I was treated with reagent for 3 min for SDS–PAGE with or without β-ME. After SDS–PAGE, the gel was stained with Coomassie brilliant blue R-250. Lanes 1 and 4, molecular markers (106 K, phosphorylase B; 98 K, bovine serum albumin; 50 K, ovalbumin; 36 K carbonic anhydrase); lane 2, without β-ME treatment; and lane 3, with β-ME treatment.

    Journal: Journal of Biochemistry

    Article Title: Disulphide linkage in mouse ST6Gal-I: determination of linkage positions and mutant analysis

    doi: 10.1093/jb/mvr133

    Figure Lengend Snippet: SDS–PAGE analysis of released mouse ST6Gal-I with or without β-mercaptoethanol (β-ME) treatment. Twenty microgram of released mouse ST6Gal-I was treated with reagent for 3 min for SDS–PAGE with or without β-ME. After SDS–PAGE, the gel was stained with Coomassie brilliant blue R-250. Lanes 1 and 4, molecular markers (106 K, phosphorylase B; 98 K, bovine serum albumin; 50 K, ovalbumin; 36 K carbonic anhydrase); lane 2, without β-ME treatment; and lane 3, with β-ME treatment.

    Article Snippet: The gel was stained with 0.2% Coomassie brilliant blue R-250 in 50% methanol and 7% acetic acid, or the gel-separated peptides were electroblotted onto a polyvinylidene difluoride membrane (Millipore, MA, USA) as described previously ( ).

    Techniques: SDS Page, Staining

    Fig. 1. TbSP1 identification. ( A ) Equal amounts of SLM-released protein were subjected to SDS–PAGE and stained with Coomassie Blue R-250. Days of in vitro culture (d) and the migration positions of molecular mass markers are indicated at the top and at the left, respectively. The migration position (p23) and the N-terminal sequence (p23/20) of the TbSP1 polypeptide are shown on the right. ( B ) Genomic DNA digested with Eco RI (lane 1), Bam HI (lane 2) or Hin dIII (lane 3) was probed with the TbSP1 cDNA. The migration positions of DNA size markers are indicated on the left. ( C ) Balanced amounts of total RNA extracted from synthetic solid (SSM) or liquid (SLM) mycelial cultures were gel fractionated and probed with the 32 P-labeled TbSP1 cDNA. The migration positions and the amounts of the 28S and 18S rRNAs, utilized as internal references, are shown on the right and in the lower panel, respectively.

    Journal: The EMBO Journal

    Article Title: A nutrient-regulated, dual localization phospholipase A2 in the symbiotic fungus Tuber borchii

    doi: 10.1093/emboj/20.18.5079

    Figure Lengend Snippet: Fig. 1. TbSP1 identification. ( A ) Equal amounts of SLM-released protein were subjected to SDS–PAGE and stained with Coomassie Blue R-250. Days of in vitro culture (d) and the migration positions of molecular mass markers are indicated at the top and at the left, respectively. The migration position (p23) and the N-terminal sequence (p23/20) of the TbSP1 polypeptide are shown on the right. ( B ) Genomic DNA digested with Eco RI (lane 1), Bam HI (lane 2) or Hin dIII (lane 3) was probed with the TbSP1 cDNA. The migration positions of DNA size markers are indicated on the left. ( C ) Balanced amounts of total RNA extracted from synthetic solid (SSM) or liquid (SLM) mycelial cultures were gel fractionated and probed with the 32 P-labeled TbSP1 cDNA. The migration positions and the amounts of the 28S and 18S rRNAs, utilized as internal references, are shown on the right and in the lower panel, respectively.

    Article Snippet: Following electrophoresis and Coomassie Blue R-250 staining, the gel was electroblotted onto Immobilon-PSQ (Millipore) and the membrane area containing the 23 kDa TbSP1 protein was cut out and subjected to N-terminal amino acid sequence analysis using a blot cartridge device and a Procise 494A protein sequencer (Applied Biosystems).

    Techniques: SDS Page, Staining, In Vitro, Migration, Sequencing, Labeling